Figure 8

Impact of IRF3 knockout and miR-122 supplementation on HCV replication. (a) Efficient knockout (K/O) of IRF3 using a CRISPR-CAS9 system. Efficient K/O of IRF3 was confirmed by western blotting. Full-length gels and blots before cropping are shown in Supplementary Fig. S6. (b) Effect of IRF3 K/O on HCV replication. H77S.3/GLuc2A and N2/GLuc2A RNAs were transfected into KH-control (CNT) and KH-IRF3 K/O cells, as well as Huh-7.5-CNT and Huh-7.5-IRF3 K/O cells. Secreted GLuc activity was determined. Differences in the means of relative GLuc activity between CNT and IRF3 K/O cells at each time point were analysed with Student’s t test. (c) Effect of IRF3 K/O and miR-122 supplementation on HCV replication. Control miRNA and miR-122, were transfected into Huh-7.5/KH-CNT cells and Huh-7.5/KH-IRF3 K/O cells, and the medium was replaced at 6 h and then every 24 h. GLuc activity at 48 h after transfection was determined and normalised to that at 48 h of control miRNA-supplemented KH-CNT cells, which was set to 1.